Objective:To solve the symptoms of achalasia complicated with osteogensis imperfecta through peroral endoscopic myotomy(POEM).Methods: The 21-year-old male patient,being diagnosed as achalasia complicated with osteogensis imperfecta,underwent POEM under general anesthesia after preoperative assessment of achalasia(Eckardt score of 7).The mucosal incision was made at a distance of 28 cm from the incisor,then the gastroscope entered the submucosal layer and tunnel in it,and myotomy was performed about 29 cm from the incisor extending 2 cm into the cardia.Electrocoagulation was performed to stop bleeding and the mucosal incision was then closed.Results:The operation was completed successfully in about 28 minutes.The stomach tube was removed 30 h after surgery.The patient was discharged in a stable condition on the fourth postoperative day.The follow-up result showed no recurrence in symptom with Eckardt score of 0 two weeks after operation.Conclusions:POEM is a feasible and effective way for achalasia of cardia complicated with osteogenesis imperfecta.

OBJECTIVETo optimize the process of tomato genetic transformation, screening and seed selection using multiepitope antigenic gene (MAG) and truncated major surface antigen 1 (tSAG1) of Toxoplasma gondii as the target insert genes.

METHODSTomato high-frequency regeneration system was optimized with different choices of media and explants. The genetic transformation procedure was optimized using different tomato cultivars, explants, culture temperatures, media and acetosyringone (AS) supplements. Three concentrations of kanamycin were utilized for resistant selection of the transgenic candidate roots. The selected lines were trained, transplanted to soil and grown in a greenhouse till maturity. Sterile seeding using kanamycin-incorporated medium was conducted for screening transgenic tomato generations.

RESULTS AND CONCLUSIONCotyledons were better than hypocotyls as the regeneration explants. The regeneration rate of cotyledons reached 98% (59/60) using the optimized regeneration medium ZB3. The culture medium and temperature were the key factors for tomato transgenic shoot induction. The number of transgenic buds increased significantly at the appropriate temperature condition (23-/+1 degrees celsius;), and AS of 100 micromol/L in the medium before inoculation also significantly raised transformation rate. The budding rate of Zhongshu No.5 cotyledons was 35% (28/81) using the medium ZB2 under (23-/+1) degrees celsius;. Kanamycin at 80 mg/L was optimal for transgenic plantlet rooting selection with the rooting rate of 48% (31/65). 117 transgenic lines were obtained. Non-transgenic tomato plant growth, especially the root and elongation, was inhibited obviously with kanamycin at 100 mg/L or above, and the roots became purple and lacked lateral roots. The transgenic tomato seeds could be selected effectively with kanamycin at 150 mg/L.

Animals ; Antigens, Protozoan ; genetics ; Drug Resistance ; Kanamycin ; pharmacology ; Lycopersicon esculentum ; drug effects ; genetics ; Plants, Genetically Modified ; drug effects ; genetics ; Protozoan Proteins ; genetics ; Seeds ; drug effects ; genetics ; Toxoplasma ; genetics ; Transformation, Genetic

OBJECTIVETo construct the plasmid reference molecules for detection of transgenic tomato line Zeneca B,Da,F using quantitative real-time PCR(qPCR).

METHODSThree plasmid reference molecules pEasy-T3-APX, pEasy-T3-16A and pEasy-T3-16S were cloned based on reverse genetics, which contain the target fragments of tomato endogenous reference gene apx (ERG-apx), gene-specific sequence of pg(GS-pg) and construct-specific sequence of vectors pJR16S/pJR16A (CS-16S/CS-16A) of Zeneca B,Da,F, respectively. Primers and Taqman probes were designed by Beacon Designer 7.5.The specificity, sensitivity, reproducibility and the limit of detection(LOD) of the qualitative and quantitative PCR system based on the plasmid reference molecules were evaluated. PicoGreen was used to measure the DNA concentration of the plasmid reference molecules. Two sets of samples containing 1% or 0.1% (w/w) pEasy-T3-16A or pEasy-T3-16S mixed with pEasy-T3- APX as background DNA were prepared for evaluating the efficacy of the qPCR system.

RESULTSThe target fragments for qPCR detection were anchored, ERG-apx 108 bp, GS-pg 108 bp , CS-16S 109 bp and CS-16A 102 bp. The three plasmid reference molecules were confirmed at the expected sizes by restriction enzyme digestion. The qPCR results showed that the RSD of reproducibility were 0.2% to1.5%, LOD was 25 copies, R2 values for these standard curves were 0.994 ~0.998 and amplification efficiencies were 93.3%~102.4%.The bias between the test and true values of two sets of mixed samples ranged from -9.3% to 14.7% after adjusting by conversion factors(Cf).

CONCLUSIONThe plasmid reference molecules and qPCR system for qualitative and quantitative detection of transgenic tomato line Zeneca B,Da,F have been established successfully.

Base Sequence ; DNA, Plant ; genetics ; Lycopersicon esculentum ; genetics ; Plants, Genetically Modified ; genetics ; Plasmids ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVE: To evaluate the effectiveness of endoscopic submucosal dissection(ESD) in treating colorectal polyps with diameter≥2.5 cm. METHODS: The clinical data of 567 cases of colorectal polyps with diameter≥2.5 cmtreated by ESD at Endoscopy Center,Zhongshan Hospital,Fudan University between January 2007 and November 2016 were analyzed retrospectively. The data included clinical and pathological characteristics, complications and follow-up.RESULTS: Of all lesions, the median diameter was 3.0 cm(2.5 to 15.0). Among them, 448 lesions(79.0%)were high grade intraepithelial neoplasia, and the other 119 lesions(21.0%)were intra-adenoma adenocarcinoma, focal cancerization or adenocarcinoma. The complete resection rate during operation was 99.1%(562/567). The en bloc resection rate was 78.3%(444/567), and the curative resection rate was 90.8%(515/567). Additional surgeries or endoscopic treatment were performed in 31 cases after ESD treatment. Postoperative bleeding occurred in 3.7%(21/567) of all cases. Penetration and electrocoagulation syndrome after ESD occurred in 1.2%(7/567) and 5.3%(30/567) of casesrespectively. The median length of follow-up was 40(12-90) months, with a local recurrence rate of 1.1%(6/536). Intraoperative complications were related to lesions ≥5.0 cm(P<0.001) and non-en bloc resection(P=0.034).Electrocoagulation syndrome was related to lesions ≥5.0 cm(P=0.004). Postponed bleeding after ESD was related to hypertension (P=0.008). Local recurrence was related to lesions ≥5.0 cm (P=0.037). CONCLUSION: Treating colorectal polyps ≥2.5 cm with ESD is safeand feasible, resulting in high rate of curative resection and an extremely low local-recurrencerate. However, polyps≥5.0 cm showld be cautionly evaluated before ESD.

OBJECTIVEThis study examined the relationship between the ultrastructural alterations of alveolar epithelial cells type II (AEC-II) and pulmonary surfactant protein A (SP-A) levels in the lung tissue of young rats with acute lung injury (ALI) in order to explore the possible mechanism of ALI.

METHODSForty-eight young Sprague-Dawley rats were randomly divided into control and ALI groups. The rats in the ALI group were intraperitoneally injected with 4 mg/kg of lipopolysaccharide (LPS) in order to induce ALI. The control subjects were injected with the same volume of normal saline. Rats were sacrificed at 24, 48 and 72 hrs after LPS or NS injection. Lung samples were obtained from the lower parts of the left lung and fixed with 2.5% glutaraldehyde for transmission electron microscope examination and for Western blot test of SP-A.

RESULTSThe microvilli of AEC-II disappeared 24 hrs after LPS injection. After 24 and 48 hrs of LPS injection, lamellar body (Lb) increased in number, enlarged in size and reduced in density, and the ring-like arrangement of Lb was present. By 48 hrs after LPS injection, giant Lb with vacuole-like deformity appeared. The contents of lung SP-A in the ALI group 24 hrs (6.52+/-0.62 vs 5.02+/-0.35; P<0.01) and 48 hrs (6.65+/-0.62 vs 5.01+/-0.36; P<0.01) after LPS injection were significantly higher than those in the control group. By 72 hrs after LPS injection, Lbs ruptured and were reduced in number. The shape of the nuclei was irregular and the border was blurred. The content of lung SP-A was greatly reduced in the ALI group 72 hrs after LPS injection compared with that in the control group (3.87+/-0.50 vs 5.22+/-0.36; P<0.01).

CONCLUSIONSThe alterations of AEC-II and lung SP-A were time-dependent in young rats with ALI induced by LPS. In the early stage of ALI, the lung SP-A content showed a compensatory increase. With the increasing injury of AEC-II cells, the secretion of SP-A presented with a decompensation and the lung SP-A content decreased. This may be one possible mechanism for the development of ARD.

Animals ; Female ; Lipopolysaccharides ; toxicity ; Male ; Microscopy, Electron ; Pulmonary Alveoli ; pathology ; ultrastructure ; Pulmonary Surfactant-Associated Protein A ; analysis ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; metabolism ; pathology

Tripolinolate A (TLA) is recently identified as a new compound from a halophyte plant Tripolium vulgare and has been shown to have significant in vitro activity against the proliferation of colorectal cancer and glioma cells. This study was designed to further investigate the effects of TLA on the proliferation of human normal cells, and the apoptosis and cell cycle in colorectal cancer cells, and the growth of tumors in the colorectal cancer-bearing animals. The data obtained from this study demonstrated that: 1) TLA had much less cytotoxicity in the human normal cells than the colorectal cancer cells; 2) TLA remarkably induced apoptosis in the human colorectal cancer cells and blocked cell cycle at G/M phase, and 3) TLA had significant anti-colorectal cancer activity in the tumor-bearing animals.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; Apoptosis ; drug effects ; Asteraceae ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Esters ; administration & dosage ; chemistry ; G2 Phase ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Phenols ; administration & dosage ; chemistry

Objective: To compare the effect of direct surgery or surgery after second-line chemotherapy for colorectal cancer patients with liver metastases who did not achieve objective remission after neoadjuvant chemotherapy. Methods: A retrospective case cohort study was used. The clinical and pathological data of 107 patients with colorectal cancer liver metastases who did not achieve objective response to neoadjuvant chemotherapy at Department of Hepatobiliary Surgery,Cancer Hospital,Chinese Academy of Medical Sciences from December 2008 to December 2016 were retrospectively collected. There were 71 males and 36 females, median age was 57 years (range: 28 to 79 years). According to the different treatment regimens after neoadjuvant chemotherapy,107 cases were divided into a direct surgery group (direct group,n=65) and an operation after receiving second-line chemotherapy group (second-line group,n=42). The propensity score matching(PSM) of the Logistic regression model was used to match the bilobar distribution of liver metastases and the number of first-line chemotherapy cycles in the two groups of patients. The caliper value was set to 0.10 and the matching ratio was 1∶2. T test, Mann-Whitney U test, χ² test or Fisher's exat test was used to analyzed the data between the tuo groups, respectively. Survival analysis design was used to investigate the difference in prognosis between the two groups of patients. Results: The follow-up time(M(IQR)) was 56.3(34.3) months (range: 2.1 to 95.0 months),and all patients were followed up. After PSM,there were 28 cases in the direct group and 42 cases in the second-line group, there were no significant differences in whether R0 resection was feasible,blood loss,blood transfusion,postoperative complications and postoperative hospital stay between the two groups (all P>0.05). The 1,3,and 5-year progression-free survival(PFS) rates of the direct group were 40.0%,16.5%,and 11.0%,and the 1,3,and 5-year overall survival(OS) rates were 98.5%,61.2%,and 41.4%,respectively, the second-line group 1,3,5 years PFS rates were 35.7%,14.3%,14.3%,1,3,5-year OS rate were 95.2%,55.1%,44.4%,respectively. The median PFS time of the direct group and the second-line group was 8.5 months and 7.5 months,respectively,and the difference was not statistically significant (P=0.826). The median OS time of the direct group and the second-line group were 33.8 months and 46.9 months,respectively. The difference was not statistically significant(P=0.646).The median PFS time of the direct group and second-line chemotherapy complete remission and partial remission group(CR/PR group) was 10.2 months and 9.1 months,respectively,and the difference was not statistically significant(P=0.669). The median OS time of the direct group and the second-line CR/PR group was 51.0 months and 46.9 months,respectively,and the difference was not statistically significant(P=0.427). The results of survival analysis suggested that major liver resection was an independent prognosis factor for PFS (HR=1.809,95%CI: 1.067 to 3.067,P=0.028) and OS(HR=2.751,95%CI: 1.317 to 5.747,P=0.007). Second-line chemotherapy was not an independent prognostic factor for PFS (HR=0.945, 95%CI:0.570 to 1.567,P=0.828) and OS (HR=0.866,95%CI: 0.468 to 1.602,P=0.646). Conclusions: There is no significant difference in the short-term outcome and long-term prognosis between direct surgery patients and second-line chemotherapy followed by surgery. Second-line chemotherapy is not an independent prognostic factor for colorectal cancer liver metastases patients who fail to achieve objective response after neoadjuvant chemotherapy.
Antineoplastic Combined Chemotherapy Protocols/therapeutic use* ; Cohort Studies ; Colorectal Neoplasms/pathology* ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms/secondary* ; Male ; Middle Aged ; Neoadjuvant Therapy ; Prognosis ; Retrospective Studies